title: Identification of amino acid residues responsible for the binding to anti-FLAG™ M2 antibody using a phage display combinatorial peptide library.
abstract: FLAG, a short hydrophilic peptide consisting of eight amino acids (DYKDDDDK), has been widely used as a fusion tag for the purification and detection of a wide variety of recombinant proteins. One of the monoclonal antibodies against this peptide, anti-FLAG M2, recognises a FLAG peptide sequence at the N terminus, Met-N terminus, C terminus, or internal site of a fusion protein and has been extremely useful for the detection, identification, and purification of recombinant proteins. Nevertheless, detailed binding specificity of anti-FLAG M2 has yet to be determined. In the current study, a phage display combinatorial peptide library was used to determine that the motif DYKxxD encompasses the critical amino acid residues responsible for the binding of FLAG peptide to this antibody. This study demonstrates the utility of phage display technology and helps to elucidate the mode of action of this detection system.
authors.affiliation: Phage Display Biotechnology Research Unit, Molecular Biotechnology Laboratory, School of Biotechnology, Institute of Agricultural Technology, Suranaree University of Technology, Nakhon Ratchasima, 30000, Thailand.
authors.forename: Witsanu
authors.initials: W
authors.lastname: Srila
authors.forename: Montarop
authors.initials: M
authors.lastname: Yamabhai
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D061505:Cell Surface Display Techniques; D002846:Chromatography, Affinity; D000939:Epitopes / Q000737:chemistry; D051379:Mice; D009842:Oligopeptides / Q000276:immunology*; D019151:Peptide Library; D011993:Recombinant Fusion Proteins / Q000276:immunology; D011994:Recombinant Proteins / Q000276:immunology
chemical_list: D000911:Antibodies, Monoclonal; D000939:Epitopes; D009842:Oligopeptides; D019151:Peptide Library; D011993:Recombinant Fusion Proteins; D011994:Recombinant Proteins; C085706:FLAG peptide